이학범 (고려대학교)

한빛사논문

조회289

이학범Hakbeom Lee

고려대학교

Biosens. Bioelectron., Feb 21 2024, 252 S0956-5663(24)00150-7 | https://doi.org/10.1016/j.bios.2024.116145 Enhanced selective discrimination of point-mutated viral RNA through false amplification regulatory direct insertion in rolling circle amplification

Hakbeom Lee ^a,1, Juneseok You ^b,1, Hansol Lee ^e, Woojoo Kim ^f, Kuewhan Jang ^c, Jinsung Park^d, Sungsoo Na ^a

^aDepartment of Mechanical Engineering, Korea University, Seoul, 02841, Republic of Korea

^bDepartment of Mechanical Engineering, Kumoh National Institute of Technology, Gumi, 31977, Republic of Korea

^cSchool of Mechanical Engineering, Hoseo University, Asan, 31499, Republic of Korea

^dDepartment of Biomechatronics Engineering, Sungkyunkwan University (SKKU), 2066 Seobu-ro, Suwon, 16419, Republic of Korea

^eAsia Pacific Influenza Institute, Korea University College of Medicine, Seoul, Republic of Korea

^fDivision of Infectious Diseases, Department of Internal Medicine, Korea University College of Medicine, Seoul, Republic of Korea

¹These authors (H. Lee and J. You) contributed equally to this work.

Corresponding authors: Kuewhan Jang, Jinsung Park, Sungsoo Na

Abstract

Coronaviruses are single-stranded RNA viruses with high mutation rates. Although a diagnostic method for coronaviruses has been developed, variants appear rapidly. Low test accuracy owing to single-point mutations is one of the main factors in the failure to prevent the early spread of coronavirus infection. Although reverse transcription-quantitative polymerase chain reaction can detect coronavirus infection, it cannot exclude the possibility of false positives, and an additional multiplexing kit is needed to discriminate single nucleotide polymorphism (SNP) variants. Therefore, in this study, we introduced a new nucleic acid amplification method to determine whether an infected person has a SNP mutation using a lateral flow assay (LFA) as a point-of-care test. Unlike traditional DNA amplification methods, direct insertion into rolling circle amplification amplifies the target genes without false amplification. After SNP-selective nucleic acid amplification, nuclease enzymes are used to make double-stranded DNA fragments that the LFA can detect, where specific mismatched DNA is found and cleaved to show different signals when a SNP-type is present. Therefore, wild- and SNP-type variants can be selectively detected. In this study, the limit of detection was 400 aM for viral RNA, and we successfully identified a dominant SNP variant selectively. Clinical tests were also conducted.

논문정보

형식Research article
게재일2024년 02월 (BRIC 등록일 2024-02-29)
연구진국내 연구진
분야 바이오·의료융합 > 바이오센싱 및 나노바이오물질

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